不同疗程高压氧治疗对脑出血大鼠血肿周围AQP4和SOD表达的影响

Effects of different hyperbaric oxygen treatment courses on perihematomal edema and expression of aquaporin-4 and superoxide dismutase in rats with intracerebral hemorrhage

目的:探究不同疗程高压氧(HBO)治疗对脑出血(ICH)大鼠血肿周围水通道蛋白(AQP4)及超氧化物歧化酶(SOD)表达的影响。方法:将52只雄性SD大鼠随机分为3组,正常组(12只)、脑出血组(20只)和高压氧治疗组(20只),脑出血模型采用胶原酶诱导法制备。各组大鼠根据不同高压氧治疗疗程再分为4个亚组:1周组、2周组、3周组和4周组,正常组每组3只大鼠,脑出血组和高压氧治疗组每组5只大鼠。高压氧治疗组于脑出血术后6h对模型制备成功的大鼠给予压力为2.0ATA,稳压吸氧时间60min的HBO治疗,1次/d,每周7次。正常组和脑出血组不予任何干预措施,所有大鼠饲养在正常环境中,各组大鼠分别在对应治疗周期结束后采用断头取脑法保留血肿周围脑组织留取标本。采用干湿比重法测定脑组织的水含量,免疫组化法测定AQP4的表达,黄嘌呤氧化酶法测定SOD的活力。结果:(1)脑组织的水含量:脑出血组及高压氧治疗组脑组织的水含量在1—3周均高于正常组(P<0.05);高压氧治疗组较脑出血组脑组织的水含量均减少(P<0.05);高压氧治疗组间1周、3周、4周脑组织水含量均低于2周(P<0.05),该三组之间差异不明显(P>0.05)。(2)脑组织AQP4的表达:脑出血组及高压氧治疗组脑组织AQP4表达在1—3周均高于正常组(P<0.05);高压氧治疗组较脑出血组1—3周脑组织AQP4表达均减少(P<0.05);高压氧治疗组间第1周及第3周与第2周相比表达较低(P<0.05)。(3)脑组织SOD的表达:脑出血组及高压氧治疗组SOD表达在1—3周均高于正常组(P<0.05),高压氧治疗组较脑出血组相比SOD表达均有增加(P<0.05);高压氧治疗组间SOD表达1周、2周、3周均高于4周(P<0.05),该3组之间相比无差异(P>0.05)。结论:高压氧治疗可下调AQP4表达减少血肿周围脑组织水含量,上调SOD表达,且连续高压氧治疗4周优于其他疗程。

Objective： To explore the effects of different courses of hyperbaric oxygen （HBO） treatment on the expression of water channel protein （aquaporin-4, AQP4） and superoxide dismutase （SOD） in the rats with intracerebral hemorrhage （ICH）. Method： A total of 52 male SD rats were randomly divided into 3 groups, normal group （12 rats）, intracerebral hemorrhage group （20 rats） and hyperbaric oxygen therapy group （20 rats）.The cerebral hemorrhage model was induced by collagenase. The rats according to different hyperbaric oxygen treatment were further divided into 4 subgroups： 1 week group, 2 weeks group, 3 weeks group and 4 weeks group with 3 rats in each normal group and 5 rats in each cerebral hemorrhage group and the hyperbaric oxygen treatment group. Hyperbaric oxygen therapy was given to the rats in 6h after the model was successfully prepared with the pressure 2.0ATA, stabilized oxygen time HBO 60min, 1 times /d, 7 times a week. Normal group and cerebral hemorrhage group were not given any intervention and all rats were kept in normal environment. The rats in each group were cut off the head to retain brain tissue around hematoma. The water content of brain tissue was measured by dry wet weight method. The expression of AQP4 was detected by immunohistochemical staining and SOD was determined by xanthine oxidase activity. The results of the experiment were processed by statistical analysis with SPSS19.0 software. Result： ①Brain tissue water content： the water content of the brain tissue in cerebral hemorrhage group and hyperbaric oxygen treatment group was higher than that in the normal group in 1—3 weeks（P＜0.05）. Moreover, the water content in the hyperbaric oxygen treatment group was lower than that in the cerebral hemorrhage group（P＜0.05）. After 1 week, 3 weeks and 4 weeks hyperbaric oxygen treatment, the water content were all lower than that after 2 weeks hyperbaric oxygen treatment（P＜0.05）②Expression of AQP4 in brain tissues： The expression of AQP4 in the cerebral hemorrhage group and the hyperbaric oxygen treatment group was higher than that in the normal group in 1—3 weeks（P＜0.05），The expression of AQP4 in the hyperbaric oxygen treatment group was lower than that in the cerebral hemorrhage group in 1—3 weeks （P＜0.05）. Compared with the 2 weeks hyperbaric oxygen treatment group, the level of AQP4 was lower than that in the 1 week and 3 weeks hyperbaric oxygen treatment group（P＜0.05）.③Expression of SOD in brain tissues： The expression of SOD in the cerebral hemorrhage group and hyperbaric oxygen therapy group was higher than that in the normal group at 1—3 weeks（P＜0.05）. Compared with the cerebral hemorrhage group, the expression of SOD in hyperbaric oxygen therapy group increased compared with that in the treatment group（P〈0.05）. The SOD expression in 1 week, 2weeks, 3 weeks hyperbaric oxygen treatment group was higher than that in 4 weeks hyperbaric oxygen treatment group （P〈0.05） and no significant difference was found between the 3 groups （P〉0.05）. Conclusion： Hyperbaric oxygen therapy can down-regulate the expression of AQP4, reduce the water content of the brain tissue around hematoma-up-regulate the expression of SOD. Moreover, the continuous hyperbaric oxygen treatment for 4 weeks is better than other treatment courses.